Site directed peptidase cleavage
AssigneeCetus Corporation
InventorsStanley N. Cohen, Annie C. Y. Chang
Life of this patent
Estimating the term…
Abstract
Method for preparing peptides free of undesired amino acids or amino acid sequences employing site specific receptors and proteases to cleave unprotected enzymatically hydrolyzable bonds.
Claims16
A method for specifically reducing the size of an extended peptide to a desired peptide product comprising:
- combining an extended peptide with at least one antibody specific for an antigenic determinant site of said extended peptide to protect an enzymatically hydrolyzable labile bond from peptidase cleavage to produce an antibody bound peptide, wherein said antibody is selected to protect a selected region of said polypeptide;
- combining said antibody bound peptide with a peptidase under conditions to provide hydrolytic cleavage of at least one exposed labile bond;
- and
- isolating said peptide product of said hydrolytic cleavage free of antibody.
A method for preparing a peptide employing hybrid DNA technology involving a DNA sequence coding for said peptide and at least one flanking amino acid resulting in an extended peptide, said method comprising:
- growing host cells containing a DNA sequence under conditions where said DNA sequence is expressed to produce an extended peptide;
- isolating said extended peptide and combining said extened peptide substantially free of said host cells with an antibody of at least one idiotype specific for an antigenic determinant site of said extended peptide to protect an enzymatically hudrolyxable labile bond from peptidase cleavage to produce an antibody bound extened peptide, wherein said antibody is selected to protect a selected region of said polypeptide;
- combining said antibody bound extened peptide with a peptidase under conditions to provide hydrolytic cleavage of at least one exposed labile bond;
- and
- isolating a peptide product of said hydrolytic cleavage free of antibody.
A method for specifically reducing the size of an extended peptide to a desired peptide product comprising:
- combining an extended peptide with an antibody of at least one idiotype specific for an antigenic determinant site to protect an enzymatically labile bond from peptidase cleavage to produce an antibody bound peptide, wherein said antibody is bound to a support;
- combining said antibody bound extened peptide with a peptidase under conditions to provide hydrolytic cleavage of at least one exposed labile bond;
- treating the resulting antibody bound peptide product with a reagent solution to free said peptide product from said antibody;
- and
- separating said peptide product from said antibody.
A kit for specifically reducing the size of an extended peptide to a desired peptide product comprising in packaged combination a multi-container unit having:
- a first container containing antibodies specific for an antigenic determinant of an extended protein wherein the antibodies protect an enzymatically hydrolyzable labile bond while leaving at least one enzymatically hudrolyzable bond exposed to peptidase hydrolysis, said antibodies being bound to a support;
- a second container containing a peptidase capable of cleaving said labile bond.
Dependencies are inferred from the claim text.